Background and purpose: Intensive care unit (ICU) nurses play an important role in nonpharmacological prevention of ventilator-associated pneumonia (VAP). The aim of this study was to assess the knowledge of nurses in ICU about non-pharmacological prevention of ventilator-associated pneumonia.Materials and methods: In a descriptive cross-sectional study, 120 ICU nurses were studied in teaching hospitals of Mazandaran University of Medical Sciences, 2015. Demographic information of nurses were recorded including age, sex, duration of being in ICU and their educational level. An authentic multiple-choice questionnaire was administered to assess the nurses' knowledge about the prevention of VAP.Results: 120 nurses including 96 females and 24 males (mean age: 32.39±5.57, mean work experience: 5.94±4.5 years) were studied. The average number of beds in the studied ICUs was 9.53±1.97 beds (Median=9). Percentage of correct answers of nurses on the oral route for intubation, frequency of ventilator circuit changes, type of airway humidifier, frequency of humidifier changes, type of suction system, frequency of change in suction systems, drainage of subglottic secretions, kinetic vs. standard beds, and patients positioning were 81.7%, 53.3%, 42.5%, 6.7%, 62.5%, 38.3%, 30%, 46.7%, and 85.8%, respectively. The average score of the participants was 4.48±1.69 out of 9.Conclusion: ICU nurses' knowledge about the type of humidifier, switching frequency of humidifier, switching frequency of suction system, subglottic secretion drainage, and the type of bed was found to be very low. These findings call for continuous training and assessment to promote the knowledge of nurses in ICUs for non-pharmaceutical prevention of VAP.